Influence of the ethylene-related signal-inhibiting octapeptide NOP-1 on postharvest ripening and quality of 'Golden Delicious' apples.

BACKGROUND: Processes extending the shelf life of climacteric fruit play an important role in terms of a sustainable global food supply. In a previous study, a synthetic octapeptide (NOP-1) was shown to inhibit the interaction between ethylene receptor (ETR) and ethylene insensitive-2 (EIN2), and in consequence delay tomato ripening. We investigated for the first time the effect of NOP-1 on inhibiting the ripening of apples ('Golden Delicious') during postharvest. RESULTS: Using purified recombinant proteins from a bacterial expression system, we demonstrate here that EIN2 also interacts tightly (Kd = 136 ± 29 nmol L-1 ) with the corresponding apple ETR MdETR1. In line with previous binding studies on tomato ETRs, the ripening-delaying peptide NOP-1 clearly binds to the purified apple ETR. An NOP-1 solution (1000 µmol L-1 ) was applied with a brush or microdispenser and compared with apples treated with 1-methylcyclopropene (1-MCP) (SmartFresh™, Agrofresh) applied as gaseous treatment or untreated control fruits. NOP-1 inhibited colour development and chlorophyll degradation during shelf life. These effects were more pronounced with the brush application (surface film) than with microdroplets application (mimicking a sprayable formulation). NOP-1 did not alter ethylene release or respiration rate, whereas 1-MCP expectedly strongly suppressed both. There were no differences in quality parameters evaluated. CONCLUSION: Our study shows that NOP-1 binds to MdETR1 which results in delaying of ethylene-dependent ripening developments of skin colour and chlorophyll. Besides application methods, possible reasons for the weak effect of NOP-1 in comparison with previous tomato experiments could be different receptor affinity and penetration differences. © 2019 Society of Chemical Industry.

Suitability of fluorescence indices for the estimation of fruit maturity compounds in tomato fruits.

BACKGROUND: We examined the suitability of chlorophyll fluorescence-based indices to monitor and predict concentrations of fruit maturity compounds during tomato ripening under different growing conditions in the greenhouse. The aim of this study was to evaluate the effects of chlorophyll concentration changes on fluorescence-based indices and to exploit the relation between fluorescence and reflectance indices with the corresponding maturity compounds determined analytically. RESULTS: Fruits grown under water deficit matured faster than control fruits as recorded with fluorescence-based indices. The SFR_R index correlated well with the analytical determination of chlorophyll content, whereas the single-signal FRF_G correlated with lycopene content even if the sensor was unable to differentiate precisely between maturity stages 2 to 4. Neither the FLAV index nor the FLAV_UV index was suitable for flavonoid prediction in tomato fruits. Compared with fluorescence indices, the relation between the reflection index and pigment concentrations was lower for chlorophyll and higher for lycopene. CONCLUSION: Chlorophyll and lycopene content in tomato fruits can be estimated by means of fluorescence indices during the pre-harvest phase. Since the chlorophyll decrease during tomato ripening is the driving force affecting all fluorescence signals, the methods are not reliable for estimation of other maturity compounds in tomato fruits. © 2018 Society of Chemical Industry.

Xeromorphic traits help to maintain photosynthesis in the perhumid climate of a Taiwanese cloud forest.

Previous flux measurements in the perhumid cloud forest of northeastern Taiwan have shown efficient photosynthesis of the endemic tree species Chamaecyparis obtusa var. formosana even under foggy conditions in which leaf surface moisture would be expected. We hypothesized this to be the result of 'xeromorphic' traits of the Chamaecyparis leaves (hydrophobicity, stomatal crypts, stomatal clustering), which could prevent coverage of stomata by precipitation, fog, and condensation, thereby maintaining CO2 uptake. Here we studied the amount, distribution, and composition of moisture accumulated on Chamaecyparis leaf surfaces in situ in the cloud forest. We studied the effect of surface tension on gas penetration to stomata using optical O2 microelectrodes in the laboratory. We captured the dynamics of condensation to the leaf surfaces with an environmental scanning electron microscope (ESEM). In spite of substantial surface hydrophobicity, the mean water film thickness on branchlets under foggy conditions was 80 µm (upper surface) and 40 µm (lower surface). This amount of water could cover stomata and prevent CO2 uptake. This is avoided by the clustered arrangement of stomata within narrow clefts and the presence of Florin rings. These features keep stomatal pores free from water due to surface tension and provide efficient separation of plant and atmosphere in this perhumid environment. Air pollutants, particularly hygroscopic aerosol, may disturb this functionality by enhancing condensation and reducing the surface tension of leaf surface water.

Acclimations to light quality on plant and leaf level affect the vulnerability of pepper (Capsicum annuum L.) to water deficit.

We investigated the influence of light quality on the vulnerability of pepper plants to water deficit. For this purpose plants were cultivated either under compact fluorescence lamps (CFL) or light-emitting diodes (LED) providing similar photon fluence rates (95 µmol m(-2) s(-1)) but distinct light quality. CFL emit a wide-band spectrum with dominant peaks in the green and red spectral region, whereas LEDs offer narrow band spectra with dominant peaks at blue (445 nm) and red (665 nm) regions. After one-week acclimation to light conditions plants were exposed to water deficit by withholding irrigation; this period was followed by a one-week regeneration period and a second water deficit cycle. In general, plants grown under CFL suffered more from water deficit than plants grown under LED modules, as indicated by the impairment of the photosynthetic efficiency of PSII, resulting in less biomass accumulation compared to respective control plants. As affected by water shortage, plants grown under CFL had a stronger decrease in the electron transport rate (ETR) and more pronounced increase in heat dissipation (NPQ). The higher amount of blue light suppressed plant growth and biomass formation, and consequently reduced the water demand of plants grown under LEDs. Moreover, pepper plants exposed to high blue light underwent adjustments at chloroplast level (e.g., higher Chl a/Chl b ratio), increasing the photosynthetic performance under the LED spectrum. Differently than expected, stomatal conductance was comparable for water-deficit and control plants in both light conditions during the stress and recovery phases, indicating only minor adjustments at the stomatal level. Our results highlight the potential of the target-use of light quality to induce structural and functional acclimations improving plant performance under stress situations.

Proximal sensing of plant-pathogen interactions in spring barley with three fluorescence techniques.

In the last years fluorescence spectroscopy has come to be viewed as an essential approach in key research fields of applied plant sciences. However, the quantity and particularly the quality of information produced by different equipment might vary considerably. In this study we investigate the potential of three optical devices for the proximal sensing of plant-pathogen interactions in four genotypes of spring barley. For this purpose, the fluorescence lifetime, the image-resolved multispectral fluorescence and selected indices of a portable multiparametric fluorescence device were recorded at 3, 6, and 9 days after inoculation (dai) from healthy leaves as well as from leaves inoculated with powdery mildew (Blumeria graminis) or leaf rust (Puccinia hordei). Genotype-specific responses to pathogen infections were revealed already at 3 dai by higher fluorescence mean lifetimes in the spectral range from 410 to 560 nm in the less susceptible varieties. Noticeable pathogen-induced modifications were also revealed by the 'Blue-to-Far-Red Fluorescence Ratio' and the 'Simple Fluorescence Ratio'. Particularly in the susceptible varieties the differences became more evident in the time-course of the experiment i.e., following the pathogen development. The relevance of the blue and green fluorescence to exploit the plant-pathogen interaction was demonstrated by the multispectral fluorescence imaging system. As shown, mildewed leaves were characterized by exceptionally high blue fluorescence, contrasting the values observed in rust inoculated leaves. Further, we confirm that the intensity of green fluorescence depends on the pathogen infection and the stage of disease development; this information might allow a differentiation of both diseases. Moreover, our results demonstrate that the detection area might influence the quality of the information, although it had a minor impact only in the current study. Finally, we highlight the relevance of different excitation-emission channels to better understand and evaluate plant-physiological alterations due to pathogen infections.

Stomatal penetration by aqueous solutions--an update involving leaf surface particles.

The recent visualization of stomatal nanoparticle uptake ended a 40-yr-old paradigm. Assuming clean, hydrophobic leaf surfaces, the paradigm considered stomatal liquid water transport to be impossible as a result of water surface tension. However, real leaves are not clean, and deposited aerosols may change hydrophobicity and water surface tension. Droplets containing NaCl, NaClO(3), (NH(4))(2) SO(4), glyphosate, an organosilicone surfactant or various combinations thereof were evaporated on stomatous abaxial and astomatous adaxial surfaces of apple (Malus domestica) leaves. The effects on photosynthesis, necrosis and biomass were determined. Observed using an environmental scanning electron microscope, NaCl and NaClO(3) crystals on hydrophobic tomato (Solanum lycopersicum) cuticles underwent several humidity cycles, causing repeated deliquescence and efflorescence of the salts. All physiological parameters were more strongly affected by abaxial than adaxial treatments. Spatial expansion and dendritic crystallization of the salts occurred and cuticular hydrophobicity was decreased more rapidly by NaClO(3) than NaCl. The results confirmed the stomatal uptake of aqueous solutions. Humidity fluctuations promote the spatial expansion of salts into the stomata. The ion-specific effects point to the Hofmeister series: chaotropic ions reduce surface tension, probably contributing to the defoliant action of NaClO(3), whereas the salt spray tolerance of coastal plants is probably linked to the kosmotropic nature of chloride ions.

Boosting leaf contents of rutin and solanesol in bio-waste of Solanum lycopersicum.

In tomato production, the accruing green biomass shows promising potential as source of health-promoting compounds, such as rutin and solanesol, that are of high interest due to their medicinal properties. Naturally, they accumulate in plants growing in suboptimal growing conditions, e.g. influenced by biotic and abiotic stressors. With the aim to evaluate the potential use of tomato residues as source, we analyzed both leaf metabolites during a complete cultivation cycle, while applying single and combined stresses practically realized in greenhouse production. In the late season, contents of both metabolites were significantly enhanced by nutrient deficit in combination with 2 °C colder nights for 4 weeks and prolonged for in total 9 weeks. Particularly, higher solanesol contents were achieved by salt stress and elevated temperature after one week, even stronger when combined with drought. At harvest, stressed plants consist of less green biomass reducing the overall economic potential. However, practicable abiotic stresses should be considered as potential tool to induce the accumulation of beneficial compounds. Extracting profitable metabolites from the green biomass of the model crop tomato supports the overall goal to promote sustainable approaches in horticultural production.

Limitation of mineral supply as tool for the induction of secondary metabolites accumulation in tomato leaves.

Agricultural residues are natural sources for secondary metabolites as high value ingredients for industrial uses. The present work aims to exploit the accumulation potential of rutin and solanesol in tomato leaves following nitrogen and general mineral deficiency in a commercial-like greenhouse. Physiological responses of tomato plants were monitored non-destructively with a multiparametric fluorescence sensor, and biochemical parameters were determined by means of HPLC analysis. Nitrogen and general mineral limitation led to an accumulation of rutin in young tomato leaves while solanesol concentration was higher in mature leaves. In young leaves, the fluorescence indices SFR_R and NBI_G showed lower values compared to control plants for both stress treatments. On the contrary, FLAV and ANTH_RG values increased during the experiment, but no differences could be recorded in mature leaves. However, correlation analysis indicates, that the FLAV index is not a reliable tool to estimate the concentration of rutin and solanesol tomato leaves. To monitor fruit yield/quality as primary objective of tomato production, fruits showing symptoms of blossom end rot (BER) were counted before and after stress treatments. BER was determined more frequently for plants grown under a general mineral deficiency, concluding that a practical applicability at the end of fruit production is advisable. Our results indicate that by-products from Solanaceae plants are promising resources for valuable bioactive leaf compounds. To achieve the highest concentrations, the seasonal variation, the optimal environmental conditions, the concentrations in different plant organs and varieties as well as different production systems are of high interest for commercial implementation.

Agricultural adjuvants may impair leaf transpiration and photosynthetic activity.

Adjuvants such as surfactants are commonly incorporated into agrochemical formulations to enhance the biological efficiency of foliar sprays by improving the wetting behavior of the spray and/or the penetration of the active ingredients into the leaf tissues. Penetration accelerating adjuvants are known to increase the cuticular permeability and may alter the cuticular barrier to water loss. However, none or very little emphasis has been given to the impacts of adjuvants on crop water balance or drought tolerance, a very important factor affecting crop performance under water scarcity. Two model crops with strongly varying leaf traits, kohlrabi (Brassica oleracea) and apple (Malus domestica) seedlings were grown in controlled environments. Three adjuvants with varying solubility in the cuticle, i.e. octanol-water partition coefficients (logKow) were selected: rapeseed methyl ester (RME) and the surfactants alkyl polyglycoside (APG) and polyoxyethylated tallow amine (POEA). The higher the logKow of the adjuvant, the stronger was the increase of minimum epidermal conductance (gmin, an essential parameter describing plant drought tolerance). However, such effects depended on the physio-chemical properties of the leaf surface. In comparison to kohlrabi, the adjuvant effects on gmin of apple leaves were relatively weak. The increase of gmin was associated with a decrease in contact angle and with an alteration of the wax microstructure. Furthermore, POEA affected photochemical efficiency of kohlrabi leaves. Some adjuvants could have a temporal influence on transpirational water loss and gmin. At repeated applications, they might alter the effective water use and possibly reduce drought tolerance of some horticultural crops.

Novel Protein-Protein Inhibitor Based Approach to Control Plant Ethylene Responses: Synthetic Peptides for Ripening Control.

Ethylene signaling is decisive for many plant developmental processes. Among these, control of senescence, abscission and fruit ripening are of fundamental relevance for global agriculture. Consequently, detailed knowledge of the signaling network along with the molecular processes of signal perception and transfer are expected to have high impact on future food production and agriculture. Recent advances in ethylene research have demonstrated that signaling of the plant hormone critically depends on the interaction of the ethylene receptor family with the NRAMP-like membrane protein ETHYLENE INSENSITIVE 2 (EIN2) at the ER membrane, phosphorylation-dependent proteolytic processing of ER-localized EIN2 and subsequent translocation of the cleaved EIN2 C-terminal polypeptide (EIN2-CEND) to the nucleus. EIN2 nuclear transport, but also interaction with the receptors sensing the ethylene signal, both, depend on a nuclear localization signal (NLS) located at the EIN2 C-terminus. Loss of the tight interaction between receptors and EIN2 affects ethylene signaling and impairs plant ethylene responses. Synthetic peptides derived from the NLS sequence interfere with the EIN2-receptor interaction and have utility in controlling plant ethylene responses such as ripening. Here, we report that a synthetic peptide (NOP-1) corresponding to the NLS motif of Arabidopsis EIN2 (aa 1262-1269) efficiently binds to tomato ethylene receptors LeETR4 and NR and delays ripening in the post-harvest phase when applied to the surface of sampled green fruits pre-harvest. In particular, degradation of chlorophylls was delayed by several days, as monitored by optical sensors and confirmed by analytical methods. Similarly, accumulation of ß-carotene and lycopene in the fruit pulp after NOP-1 application was delayed, without having impact on the total pigment concentration in the completely ripe fruits. Likewise, the peptide had no negative effects on fruit quality. Our molecular and phenotypic studies reveal that peptide biologicals could contribute to the development of a novel family of ripening inhibitors and innovative ripening control in climacteric fruit.

Leaf surface characteristics of apple seedlings, bean seedlings and kohlrabi plants and their impact on the retention and rainfastness of mancozeb.

A study was made of the influence of the upper leaf surface characteristics on the retention and rainfastness of the contact fungicide mancozeb with and without tank-mix adjuvants (RSO 5 and RSO 60) on apple seedlings, bean seedlings and kohlrabi plants. Large differences in roughness, in the amount and composition of surface waxes and in the retention and rainfastness of mancozeb were found among species. Strong correlations between roughness and total amount of surface waxes and mass of C29 alkane in the wax mass were also found. Fungicide retention was strongly, negatively correlated with surface roughness, total epicuticular wax, amount of C29 alkane and the total mass of alkanes. Rainfastness correlated strongly or very strongly with the amount of C28 alcohol and C33 alkane. The addition of a more hydrophobic (RSO 5) or a more hydrophilic (RSO 60) adjuvant to the spray solution influenced retention and rainfastness, and also altered the correlation coefficients. The present results support earlier observations which show that the success of adjuvants in enhancing the retention and rainfastness of agrochemicals depends on the characteristics of the leaf surface.

Concentration of hinokinin, phenolic acids and flavonols in leaves and stems of Hydrocotyle leucocephala is differently influenced by PAR and ecologically relevant UV-B level.

We examined the effects of ambient, non-stressing ultraviolet (UV)-B (280-315nm) level combined with different intensities of photosynthetic active radiation (PAR, 400-700nm) on the accumulation of the lignan (-)-hinokinin, in leaves and stems of Hydrocotyle leucocephala. Plants were exposed in sun simulators under almost natural irradiance and climatic conditions to one of four light regimes, i.e. two PAR intensities (906 and 516µmolm(-2)s(-1)) including or excluding UV-B radiation (0 and 0.4Wm(-2)). Besides hinokinin, we identified three chlorogenic acid isomers, one other phenolic acid, 12 quercetin, and five kaempferol derivatives in the H. leucocephala extracts. Hinokinin was most abundant in the stems, and its accumulation was slightly enhanced under UV-B exposure. We therefore assume that hinokinin contributes to cell wall stabilization and consequently to a higher resistance of the plant to environmental factors. Quercetin derivatives increasingly accumulated under UV-B and high PAR exposure at the expense of kaempferols and chlorogenic acids, which was apparently related to its ability to scavenge reactive oxygen species. In general, the concentration of the constituents depended on the plant organ, the leaf age, the light regimes, and the duration of exposure. The distribution pattern of the compounds within the examined organs was not influenced by the treatments. Based on the chemical composition of the extracts a principal component analysis (PCA) enabled a clear separation of the plant organs and harvesting dates. Younger leaves mostly contained higher phenylpropanoid concentrations than older leaves. Nevertheless, more pronounced effects of the light regimes were detected in older leaves. As assessed, in many cases the individual compounds responded differently to the PAR/UV-B combinations, even within the same phenylpropanoid class. Since this is the first report on the influence of light conditions on the accumulation of lignans in herbaceous plants, it opens many perspectives for a more precise elucidation of all involved biochemical and molecular processes.

Assessment of photosystem II thermoluminescence as a tool to investigate the effects of dehydration and rehydration on the cyclic/chlororespiratory electron pathways in wheat and barley leaves.

Thermoluminescence emission from wheat leaves was recorded under various controlled drought stress conditions: (i) fast dehydration (few hours) of excised leaves in the dark (ii) slow dehydration (several days) obtained by withholding watering of plants under a day/night cycle (iii) overnight rehydration of the slowly dehydrated plants at a stage of severe dessication. In fast dehydrated leaves, the AG band intensity was unchanged but its position was shifted to lower temperatures, indicating an activation of cyclic and chlororespiratory pathways in darkness, without any increase of their overall electron transfer capacity. By contrast, after a slow dehydration the AG intensity was strongly increased whereas its position was almost unchanged, indicating respectively that the capacity of cyclic pathways was enhanced but that they remained inactivated in darkness. Under more severe dehydration, the AG band almost disappeared. Rewatering caused its rapid bounce significantly above the control level. No significant differences in AG emission could be found between the two drought-sensitive and drought-tolerant wheat cultivars. The afterglow thermoluminescence emission in leaves provides an additional tool to follow the increased capacity and activation of cyclic electron flow around PSI in leaves during mild, severe dehydration and after rehydration.

"Breath figures" on leaf surfaces-formation and effects of microscopic leaf wetness.

"Microscopic leaf wetness" means minute amounts of persistent liquid water on leaf surfaces which are invisible to the naked eye. The water is mainly maintained by transpired water vapor condensing onto the leaf surface and to attached leaf surface particles. With an estimated average thickness of less than 1 µm, microscopic leaf wetness is about two orders of magnitude thinner than morning dewfall. The most important physical processes which reduce the saturation vapor pressure and promote condensation are cuticular absorption and the deliquescence of hygroscopic leaf surface particles. Deliquescent salts form highly concentrated solutions. Depending on the type and concentration of the dissolved ions, the physicochemical properties of microscopic leaf wetness can be considerably different from those of pure water. Microscopic leaf wetness can form continuous thin layers on hydrophobic leaf surfaces and in specific cases can act similar to surfactants, enabling a strong potential influence on the foliar exchange of ions. Microscopic leaf wetness can also enhance the dissolution, the emission, and the reaction of specific atmospheric trace gases e.g., ammonia, SO2, or ozone, leading to a strong potential role for microscopic leaf wetness in plant/atmosphere interaction. Due to its difficult detection, there is little knowledge about the occurrence and the properties of microscopic leaf wetness. However, based on the existing evidence and on physicochemical reasoning it can be hypothesized that microscopic leaf wetness occurs on almost any plant worldwide and often permanently, and that it significantly influences the exchange processes of the leaf surface with its neighboring compartments, i.e., the plant interior and the atmosphere. The omission of microscopic water in general leaf wetness concepts has caused far-reaching, misleading conclusions in the past.

Ecologically relevant UV-B dose combined with high PAR intensity distinctly affect plant growth and accumulation of secondary metabolites in leaves of Centella asiatica L. Urban.

We investigated the effects of environmentally relevant dose of ultraviolet (UV)-B and photosynthetic active radiation (PAR) on saponin accumulation in leaves on the example of Centella asiatica L. Urban. For this purpose, plants were exposed to one of four light regimes i.e., two PAR intensities with or without UV-B radiation. The experiment was conducted in technically complex sun simulators under almost natural irradiance and climatic conditions. As observed, UV-B radiation increased herb and leaf production as well as the content of epidermal flavonols, which was monitored by non-destructive fluorescence measurements. Specific fluorescence indices also indicate an increase in the content of anthocyanins under high PAR; this increase was likewise observed for the saponin concentrations. In contrast, UV-B radiation had no distinct effects on saponin and sapogenin concentrations. Our findings suggest that besides flavonoids, also saponins were accumulated under high PAR protecting the plant from oxidative damage. Furthermore, glycosylation of sapogenins seems to be important either for the protective function and/or for compartmentalization of the compounds. Moreover, our study revealed that younger leaves contain higher amounts of saponins, while in older leaves the sapogenins were the most abundant constituents. Concluding, our results proof that ambient dose of UV-B and high PAR intensity distinctly affect the accumulation of flavonoids and saponins, enabling the plant tissue to adapt to the light conditions.

Centelloside accumulation in leaves of Centella asiatica is determined by resource partitioning between primary and secondary metabolism while influenced by supply levels of either nitrogen, phosphorus or potassium.

In the present study we aimed to investigate the relevance of either N, P or K supply for herb and leaf yield and for centelloside concentrations in Centella asiatica L. Urban leaves. In this regard, we elucidated the causal relationship between assimilation rate, leaf N, P and K concentrations, herb and leaf production, and centelloside accumulation. The experiments were conducted consecutively in a greenhouse where C. asiatica was grown in hydroponic culture and fertigated with nutrient solutions at either 0, 30, 60, 100 or 150% of the N, P or K amount in a standard Hoagland solution. In general, the increase in N, P or K supply enhanced assimilation rate and herb and leaf yield. However, exceeding specific thresholds, the high availability of one single nutrient caused lower leaf N concentrations and a decline in assimilation rate and plant growth. Irrespective of N, P and K supply, the leaf centelloside concentrations were negatively associated with herb and leaf yield, which is in accordance with the assumptions of the carbon/nutrient balance and the growth differentiation balance hypotheses. Moreover, we found strong negative correlations between saponins and leaf N concentrations, while the respective sapogenins were negatively correlated with K concentrations. Using C. asiatica as model system, our experiments reveal for the first time that the accumulation of saponins and sapogenins is affected by resource allocation between primary and secondary metabolism and that besides carbon, also nutrient availability is relevant for the regulation of the centelloside synthesis. Finally, our results highlight the huge potential of optimized and carefully controlled mineral nutrition of medicinal plants for steering the bio-production of high-quality natural products.

Effects of relative humidity and substrate on the spatial association between glyphosate and ethoxylated seed oil adjuvants in the dried deposits of sessile droplets.

BACKGROUND: In recent years, several studies have shown the impact of adjuvants on the characteristics of herbicide deposits on leaf surfaces. Until now, most studies have addressed the distribution of active ingredients (AIs), whereas few experiments have focused on the location of the adjuvants. The objective of this study was a systematic examination of the particle distribution profile of both the AI (glyphosate, Gly) and the adjuvants after the application of sessile microdroplets on hydrophobic (Teflon) and hydrophilic (glass and aluminium) model surfaces. RESULTS: The association degree (AD) was surface dependent and specific for the tested adjuvants. In general, the rather hydrophobic adjuvant RSO 5 showed decreasing AD with Gly at increasing relative humidity (RH) levels. The rather hydrophilic RSO 60 adjuvant displayed higher AD between the compounds at a higher RH. A high concentration of the adjuvant reduced the AD for both of the RSO adjuvants evaluated. CONCLUSION: The combination of surface properties, the type of adjuvant and the relative humidity determines the degree of association between Gly and the adjuvants. The present results suggest that the interaction between the AI and an adjuvant determines whether spatial separation occurs, whereas physical processes (e.g. capillary particle movement, inward and outward Marangoni flows and the evaporation rate) are decisive for the extent of the separation. Coffee-ring structures were formed exclusively with the adjuvant+Gly mixtures, whereas Gly alone formed either one big deposit or several small islands distributed within the droplet footprint.

Presymptomatic detection of powdery mildew infection in winter wheat cultivars by laser-induced fluorescence.

The sensor-based monitoring of diseases under controlled conditions establishes an objective tool that allows a better understanding of the pathogen-plant interactions in different situations. The purpose of our work was to implement the presymptomatic detection of powdery mildew on wheat leaves shortly after fungus inoculation by spectral and time-resolved laser-induced fluorescence spectroscopy. In the general scope of plant phenotyping, we hypothesized that it is possible to discriminate between wheat genotypes that are either resistant or susceptible to powdery mildew. According to our results, the presymptomatic detection of powdery mildew on wheat leaves was accomplished, irrespective of genotype, as early as one day after inoculation using the fluorescence amplitude ratio F451:F522. Similarly, the ratios F451:F522, F522:F687, and F522:F736 of the half-bandwidth are also appropriate parameters. Furthermore, in the spectral range between 410 nm and 620 nm, the mean lifetime was significantly longer in inoculated leaves than it was in control leaves. Finally, the short-term (10-12 hour) increase of the fluorescence mean lifetime at 530 nm and 560 nm following the inoculation suggests that the speed of the plant reaction might be associated to its resistance to the pathogen. Based on this information, we conclude that determinations of ultraviolet, laser-induced fluorescence intensity and lifetime are suitable approaches to presymptomatically detect powdery mildew on wheat leaves one day after inoculation.

Use of blue-green and chlorophyll fluorescence measurements for differentiation between nitrogen deficiency and pathogen infection in winter wheat.

In recent years, several sensor-based approaches have been established to early detect single plant stresses, but the challenge of discriminating between simultaneously occurring stressors still remains. Earlier studies on wheat plants strongly affected by pathogens and nitrogen deficiency indicated that chlorophyll fluorescence might be suited to distinguish between the two stressors. Nevertheless, there is lack of information on the pre-symptomatic detection of synchronized occurrence of slight N-deficiency and the early stages of pathogen infection. The usefulness of the blue, green, and yellow fluorescence signals in this context has not yet been explored. We hypothesized that differentiation between wheat plants' physiological reaction due to N-deficiency and leaf rust (Puccinia triticina) as well as N-deficiency and powdery mildew (Blumeria graminis f. sp. tritici) might be accomplished by means of UV laser-induced fluorescence spectral measurements between 370 and 620nm in addition to chlorophyll fluorescence (640-800nm). Plants were provided with either a normal or a modified Hoagland nutrient solution in order to induce a slight N deficit. Pathogen inoculation was carried out on the second fully developed leaf. Four experimental groups were evaluated: (a) N-full-supply [N+]; (b) N-deficiency [N-]; (c) N-full-supply+pathogen [N+/LR] or [N+/PM]; (d) N-deficiency+pathogen [N-/LR] or [N-/PM]. The results revealed that, in addition to the amplitude ratio of R/FR fluorescence, B/G fluorescence also facilitated reliable and robust discrimination among the four experimental groups. The discrimination among the experimental groups was accomplished as early as one and two days after inoculation for powdery mildew and leaf rust infection, respectively. During the 3days evaluation period, the differences among the treatment groups became more evident. Moreover, several other amplitude ratios and half-bandwidth ratios proved to be suited to early detect fungal infection, irrespective of the nitrogen status of the plant.

UV-induced fluorescence spectra and lifetime determination for detection of leaf rust (Puccinia triticina) in susceptible and resistant wheat (Triticum aestivum) cultivars.

In modern agriculture, the use of cultivars that are resistant against specific stresses, e.g. pathogen infections, is an integral component. Considering the great demand for a rapid and objective screening method for stress resistance of new cultivars, the question arises, whether time resolved fluorescence spectroscopy is suitable for such purposes. Amongst others, infected plants might accumulate specific compounds such as salicylic acid and phenylpropanoid compounds as key substances in plant disease resistance, whereas synthesis and accumulation may influence fluorescence parameters such as absolute intensity of single peaks, ratios between peaks and lifetime. Experiments were conducted in a controlled-environment cabinet cultivating four leaf rust susceptible and three leaf rust resistant genotypes. Fluorescence measurements were conducted using a compact fibre-optic fluorescence spectrometer with a nanosecond time-resolution. Results of experiments revealed that UV-induced measurements of spectral characteristics as well as determination of fluorescence lifetime are suited to detect leaf rust (Puccinia triticina) in wheat (Triticum aestivum L.) cultivars as early as 2 days after inoculation (dai). For this purpose several parameters such as the fluorescence (F) amplitude ratios F451/F522, F451/F687, F451/F736, F522/F687, F522/F736 as well as fluorescence mean lifetime especially at 470nm, might be used. Discrimination between resistant and susceptible cultivars to the leaf rust pathogen could be accomplished 3dai by using the ratio of fluorescence amplitude between the blue (F451nm) and red (F687nm) peak, and mean lifetime at 440, 500 and 530nm. Our results indicate that the combination of spectrally and time-resolved fluorescence could be an additional tool in plant breeding programs for an automatic and precise high-throughput system for evaluation of the pathogen resistance of new genotypes.