Photon- and Singlet-Oxygen-Induced Cis-Trans Isomerization of the Water-Soluble Carotenoid Crocin.

Studying the cis-trans isomerization process in crocin (CR), one of the few water-soluble carotenoids extracted from saffron, is important to better understand the physiological role of cis-carotenoids in vivo and their potential as antioxidants in therapeutic applications. For that, cis-trans isomerization of both methanol- and water-dissolved CR was induced by light or thermally generated singlet oxygen (1O2). The kinetics of molecular concentrations were monitored by both high-performance liquid chromatography (HPLC) and non-destructive spectrophotometric methods. These last made it possible to simultaneously follow the cis-trans isomerization, the possible bleaching of compounds and the amount of thermally generated 1O2. Our results were in accordance with a comprehensive model where the cis-trans isomerization occurs as relaxation from the triplet state of all-trans- or 13-cis-CR, whatever is the way to populate the CR triplet state, either by photon or 1O2 energy transfer. The process is much more (1.9 to 10-fold) efficient from cis to trans than vice versa. In H2O, a 1O2-induced bleaching effect on the starting CR was not negligible. However, the CR "flip-flop" isomerization reaction could still occur, suggesting that this process can represent an efficient mechanism for quenching of reactive oxygen species (ROS) in vivo, with a limited need of carotenoid regeneration.

Non-destructive fluorescence sensing for assessing microclimate, site and defoliation effects on flavonol dynamics and sugar prediction in Pinot blanc grapes.

In an era of growing international competition in modern viticulture, the study and implementation of innovative technologies to increase the production of high-quality grapes and wines are of critical importance. In this study, the non-destructive portable sensor Multiplex, based on fluorescence sensing technique, was applied to evaluate grape maturity parameters and flavonol content of the understudied Pinot blanc variety. The effects of environmental and agronomical factors on flavonol content of Pinot blanc grapes were investigated in eight vineyards characterised by different microclimatic and agronomic conditions. Furthermore, the direct impact of canopy management treatment on the flavonol dynamics of the grapes oriented in the four cardinal directions was assessed. Results highlight the positive role of moderate temperatures and direct sunlight exposure on Pinot blanc flavonol content; however, no direct vineyard-elevation effect was observed. The ability to modulate and evaluate the flavonol content in field represent crucial factors because of their potential effect on flavonoids-dependent wine characteristics, such as stability and ageing. In the present study, for the first time, two calibration curves were reported for pre- and post-veraison periods between flavonol indices and the berry skin flavonol content and a good correlation was observed between Multiplex measurement and the total polyphenolic content of grape juice. Moreover, the strong correlation between the chlorophyll index with grape juice sugar content and titratable acidity revealed the practical application of non-destructive sensors to predict the optimal harvest time for Pinot blanc grapes. In conclusion, the non-destructive fluorescence sensor Multiplex is a high-potential tool for innovative viticulture, for evaluating grape skin composition variables in white grape varieties.

Retrieving the in vivo Scopoletin Fluorescence Excitation Band Allows the Non-invasive Investigation of the Plant-Pathogen Early Events in Tobacco Leaves.

In this study, we developed and applied a new spectroscopic fluorescence method for the in vivo detection of the early events in the interaction between tobacco (Nicotiana tabacum L.) plants and pathogenic bacteria. The leaf disks were infiltrated with a bacterial suspension in sterile physiological solution (SPS), or with SPS alone as control. The virulent Pseudomonas syringae pv. tabaci strain ATCC 11528, its non-pathogenic ΔhrpA mutant, and the avirulent P. syringae pv. tomato strain DC3000 were used. At different post-infiltration time-points, the in vivo fluorescence spectra on leaf disks were acquired by a fiber bundle-spectrofluorimeter. The excitation spectra of the leaf blue emission at 460 nm, which is mainly due to the accumulation of coumarins following a bacterial infiltration, were processed by using a two-bands Gaussian fitting that enabled us to isolate the scopoletin (SCT) contribution. The pH-dependent fluorescence of SCT and scopolin (SCL), as determined by in vitro data and their intracellular localization, as determined by confocal microscopy, suggested the use of the longer wavelength excitation band at 385 nm of 460 nm emission (F385_460) to follow the metabolic evolution of SCT during the plant-bacteria interaction. It was found to be directly correlated (R 2 = 0.84) to the leaf SCT content, but not to that of SCL, determined by HPLC analysis. The technique applied to the time-course monitoring of the bacteria-plant interaction clearly showed that the amount and the timing of SCT accumulation, estimated by F385_460, was correlated with the resistance to the pathogen. As expected, this host defense response was delayed after P. syringae pv. tabaci ATCC 11528 infiltration, in comparison to P. syringae pv. tomato DC3000. Furthermore, no significant increase of F385_460 (SCT) was observed when using the non-pathogenic ΔhrpA mutant of P. syringae pv. tabaci ATCC 11528, which lacks a functional Type Three Secretion System (TTSS). Our study showed the reliability of the developed fluorimetric method for a rapid and non-invasive monitoring of bacteria-induced first events related to the metabolite-based defense response in tobacco leaves. This technique could allow a fast selection of pathogen-resistant cultivars, as well as the on-site early diagnosis of tobacco plant diseases by using suitable fluorescence sensors.

Valorization of waste cabbage leaves by postharvest photochemical treatments monitored with a non-destructive fluorescence-based sensor.

The biosynthesis of polyphenolic compounds in cabbage waste, outer green leaves of white head cabbage (Brassica oleracea L. var. capitata subvar. alba), was stimulated by postharvest irradiation with UVB lamps or sunlight. Both treatments boosted the content of kaempferol and quercetin glycosides, especially in the basal leaf zone, as determined by the HPLC analysis of leaf extracts and by a non-destructive optical sensor. The destructive analysis of samples irradiated by the sun for 6 days at the end of October 2015 in Skierniewice (Poland) showed an increase of leaf flavonols by 82% with respect to controls. The treatment by a broadband UVB fluorescent lamp, with irradiance of 0.38 W m-2 in the 290-315 nm range (and 0.59 W m-2 in the UVA region) for 12 h per day at 17 °C along with a white light of about 20 µmol m-2 s-1, produced a flavonols increase of 58% with respect to controls. The kinetics of flavonols accumulation in response to the photochemical treatments was monitored with the FLAV non-destructive index. The initial FLAV rate under the sun was proportional to the daily radiation doses with a better correlation for the sun global irradiance (R2 = 0.973), followed by the UVA (R2 = 0.965) and UVB (R2 = 0.899) irradiance. The sunlight turned out to be more efficient than the UVB lamp in increasing the flavonols level of waste leaves, because of a significant role played by UVA and visible solar radiation in the regulation of the flavonoid accumulation in cabbage. The FLAV index increase induced on the adaxial leaf side was accompanied by a lower but still significant FLAV increase on the unirradiated abaxial side, likely due to a systemic signaling by mean of the long-distance movement of macromolecules. Our present investigation provides useful data for the optimization of postharvest photochemical protocols of cabbage waste valorization. It can represent a novel and alternative tool of vegetable waste management for the recovery of beneficial phytochemicals.

Application of a Combined Transmittance/Fluorescence Leaf Clip Sensor for the Nondestructive Determination of Nitrogen Status in White Cabbage Plants.

The correct fertilization of vegetable crops is commonly determined on the basis of soil and plant costly destructive analyses, demanding more sustainable non-invasive optical detection. Here, we tested the ability of the combined transmittance/fluorescence leaf clip Dualex device for determining the nitrogen (N) status of cabbage plants. Fully developed leaves from plants grown under different N rates of 0; 100; 200; 300 kg N ha-1 in 2018 and 2019 were measured in the field by the Dualex sensor twice a year in July and October. The chlorophyll (Chl) and nitrogen (nitrogen balance index, NBI) indices and the flavonols (Flav) index of the sensor were positively and negatively correlated to leaf nitrogen, respectively. Merging the two-years data, the NBI versus leaf N correlation was less point dispersed in October than July (R2 = 0.76 and 0.64, respectively). NBI was also correlated to cabbage yield, better in July than October. Our results showed that the multiparametric Dualex device can be used as precision agriculture tool for the early prediction of plant N and cabbage yield with economic advantage for the growers and reduced environmental contamination due to nitrate leaching.

Unveiling the shade nature of cyanic leaves: A view from the "blue absorbing side" of anthocyanins.

Anthocyanins have long been suggested as having great potential in offering photoprotection to plants facing high light irradiance. Nonetheless, their effective ability in protecting the photosynthetic apparatus from supernumerary photons has been questioned by some authors, based upon the inexact belief that anthocyanins almost exclusively absorb green photons, which are poorly absorbed by chlorophylls. Here we focus on the blue light absorbing features of anthocyanins, a neglected issue in anthocyanin research. Anthocyanins effectively absorb blue photons: the absorbance of blue relative to green photons increases from tri- to mono-hydroxy B-ring substituted structures, reaching up to 50% of green photons absorption. We offer a comprehensive picture of the molecular events activated by low blue-light availability, extending our previous analysis in purple and green basil, which we suggest to be responsible for the "shade syndrome" displayed by cyanic leaves. While purple leaves display overexpression of genes promoting chlorophyll biosynthesis and light harvesting, in green leaves it is the genes involved in the stability/repair of photosystems that are largely overexpressed. As a corollary, this adds further support to the view of an effective photoprotective role of anthocyanins. We discuss the profound morpho-anatomical adjustments imposed by the epidermal anthocyanin shield, which reflect adjustments in light harvesting capacity under imposed shade and make complex the analysis of the photosynthetic performance of cyanic versus acyanic leaves.

Are Flavonoids Effective Antioxidants in Plants? Twenty Years of Our Investigation.

Whether flavonoids play significant antioxidant roles in plants challenged by photooxidative stress of different origin has been largely debated over the last few decades. A critical review of the pertinent literature and our experimentation as well, based on a free-of-scale approach, support an important antioxidant function served by flavonoids in plants exposed to a wide range of environmental stressors, the significance of which increases with the severity of stress. On the other side, some questions need conclusive answers when the putative antioxidant functions of plant flavonoids are examined at the level of both the whole-cell and cellular organelles. This partly depends upon a conclusive, robust, and unbiased definition of "a plant antioxidant", which is still missing, and the need of considering the subcellular re-organization that occurs in plant cells in response to severe stress conditions. This likely makes our deterministic-based approach unsuitable to unveil the relevance of flavonoids as antioxidants in extremely complex biological systems, such as a plant cell exposed to an ever-changing stressful environment. This still poses open questions about how to measure the occurred antioxidant action of flavonoids. Our reasoning also evidences the need of contemporarily evaluating the changes in key primary and secondary components of the antioxidant defense network imposed by stress events of increasing severity to properly estimate the relevance of the antioxidant functions of flavonoids in an in planta situation. In turn, this calls for an in-depth analysis of the sub-cellular distribution of primary and secondary antioxidants to solve this still intricate matter.

Short-Term Pre-Harvest UV-B Supplement Enhances the Polyphenol Content and Antioxidant Capacity of Ocimum basilicum Leaves during Storage.

Ocimum basilicum (basil) leaves are rich in polyphenols, conferring them a high antioxidant activity. The application of UV-B can be used to maintain the post-harvest nutraceutical quality of basil leaves. We aimed to investigate the effects of pre-harvest UV-B application on polyphenolic and pigment contents, antioxidant capacity, and the visual quality of basil stored leaves. We also evaluated the applicability of the non-invasive Dualex® for monitoring the accumulation of leaf epidermal phenolics (Flav Index). After exposing plants to white light (control) and to supplemental UV-B radiation for 4 d, the leaves were harvested and stored for 7d (TS7). The UV-B leaves showed both a higher phenolic content and antioxidant capacity than the controls at TS7. In addition, the correlations between the Flav Index and phenolic content demonstrated that Dualex® can reliably assess the content of epidermal phenolics, thus confirming its promising utilization as a non-destructive method for monitoring the phytochemical quality of O. basilicum leaves. In conclusion, a pre-harvesting UV-B application may be a tool for enhancing the content of polyphenols and the antioxidant potential of basil stored leaves without detrimental effects on their visual quality. These results are important considering the nutraceutical value of this plant and its wide commercial distribution.

Fluorescence-Sensor Mapping for the in Vineyard Non-Destructive Assessment of Crimson Seedless Table Grape Quality.

Non-destructive tools for the in situ evaluation of vine fruit quality and vineyard management can improve the market value of table grape. We proposed a new approach based on a portable fluorescence sensor to map the ripening level of Crimson Seedless table grape in five different plots in the East, Central-North and South of the Macedonia Region of Greece. The sensor provided indices of ripening and color such as SFRR and ANTHRG correlated to the chlorophyll and anthocyanin berry contents, respectively. The mean ANTHRG index was significantly different among all the plots examined due to the occurrence of different environmental conditions and/or asynchronous ripening processes. The indices presented moderate, poor in some cases, spatial variability, probably due to a significant vine-to-vine, intra-vine and intra-bunch variability. The cluster analysis was applied to the plot with the most evident spatial structure (at Kilkis). Krigged maps of the SFRR, ANTHRG and yield were classified by k-means clustering in two-zones that differed significantly in their mean values. ANTHRG and SFRR were inversely correlated over 64% of the plot. SFRR appeared to be a potential useful proxy of yield since it was directly correlated to yield over 66% of the plot. The grape color (ANTHRG) was slightly higher over the low-yield zones with respect to the high-yield zones. Our study showed that the combination of anthocyanins and chlorophyll indices detected in the field on Crimson Seedless table grape by a portable fluorescence sensor can help in defining the best harvest time and the best areas for harvesting.

Valorization of Tomato Surplus and Waste Fractions: A Case Study Using Norway, Belgium, Poland, and Turkey as Examples.

There is a large potential in Europe for valorization in the vegetable food supply chain. For example, there is occasionally overproduction of tomatoes for fresh consumption, and a fraction of the production is unsuited for fresh consumption sale (unacceptable color, shape, maturity, lesions, etc.). In countries where the facilities and infrastructure for tomato processing is lacking, these tomatoes are normally destroyed, used as landfilling or animal feed, and represent an economic loss for producers and negative environmental impact. Likewise, there is also a potential in the tomato processing industry to valorize side streams and reduce waste. The present paper provides an overview of tomato production in Europe and the strategies employed for processing and valorization of tomato side streams and waste fractions. Special emphasis is put on the four tomato-producing countries Norway, Belgium, Poland, and Turkey. These countries are very different regards for example their climatic preconditions for tomato production and volumes produced, and represent the extremes among European tomato producing countries. Postharvest treatments and applications for optimized harvest time and improved storage for premium raw material quality are discussed, as well as novel, sustainable processing technologies for minimum waste and side stream valorization. Preservation and enrichment of lycopene, the primary health promoting agent and sales argument, is reviewed in detail. The European volume of tomato postharvest wastage is estimated at >3 million metric tons per year. Together, the optimization of harvesting time and preprocessing storage conditions and sustainable food processing technologies, coupled with stabilization and valorization of processing by-products and side streams, can significantly contribute to the valorization of this underutilized biomass.

Comparison of lycopene changes between open-field processing and fresh market tomatoes during ripening and post-harvest storage by using a non-destructive reflectance sensor.

BACKGROUND: Accumulation and stability of tomato lycopene markedly depends on the cultivar, plant growing and storage conditions. To estimate lycopene in open-field cultivated processing and fresh market tomatoes, we used a calibrated spectral reflectance portable sensor. RESULTS: Lycopene accumulation in fruits attached to the plant, starting from the Green ripening stage, followed a sigmoidal function. It was faster and reached higher levels in processing (cv. Calista) than fresh market (cv. Volna) tomatoes (90 and 62 mg kg-1 fresh weight, respectively). During storage at 12, 20 and 25 °C, Red tomatoes retained about 90% of harvest lycopene for three weeks. Pink tomatoes increased lycopene during the first week of storage, but never reached the lycopene values of Red tomatoes ripened on the vine. Storability at 12 °C retaining the highest quality in red tomatoes was limited to 14 and 7 days for Calista and Volna cultivars, respectively. CONCLUSION: Significant differences in lycopene accumulation and stability between processing and fresh market tomatoes were established by examining with time the very same fruits by a non-destructive optical tool. It can be useful in agronomical and post-harvest physiological studies and can be of interest for producers oriented to the niche nutraceutical market. © 2018 Society of Chemical Industry.

Multiple Consequences Induced by Epidermally-Located Anthocyanins in Young, Mature and Senescent Leaves of Prunus.

Anthocyanic morphs are generally less efficient in terms of carbon gain, but, in turn, are more photoprotected than anthocyanin-less ones. To date, mature leaves of different morphs or leaves at different developmental stages within the same species have generally been compared, whereas there is a lack of knowledge regarding different stages of development of red vs. green leaves. Leaves (1-, 7-, and 13-week-old) of red- (RLP) and green-leafed (GLP) Prunus in terms of photosynthetic rate, carbon metabolism and photoprotective mechanisms were compared to test whether anthocyanin-equipped leaves perform better than anthocyanin-less leaves and whether photoprotection is the primary role of epidermally-located anthocyanins, using for the first time a recently-developed parameter of chlorophyll fluorescence (qPd). GLP leaves had a higher photosynthetic rate in 1- and 7-week-old leaves, but RLP leaves performed better at an early stage of senescence and had a longer leaf lifespan. Anthocyanins contributed to leaf photoprotection throughout the leaf development, but were tightly coordinated with carotenoids. Besides photoprotecting, we propose that epidermal anthocyanins may be principally synthetized to maintain an efficient carbon-sink strength in young and senescent leaves, thus extending the RLP leaf lifespan.

Management Zone Delineation for Winegrape Selective Harvesting Based on Fluorescence-Sensor Mapping of Grape Skin Anthocyanins.

We analyzed the potential of non-destructive optical sensing of grape skin anthocyanins for selective harvesting in precision viticulture. We measured anthocyanins by a hand-held fluorescence optical sensor on a 7 ha Sangiovese vineyard plot in central Italy. Optical indices obtained by the sensor were calibrated for the transformation in units of anthocyanins per berry mass, i.e., milligrams per gram of berry fresh weight. A full protocol for optimal data filtration, interpolation, and homogeneous zone delineation based on a very large number of optical measurements is proposed. Both the single signal-based fluorescence index (ANTHR) and the two signal ratio-based index (ANTHRG) can be used for Sangiovese grapes. Significant separations of grape-quality batches were obtained by several methods of data classification and zone delineations. Basic statistical criteria were as efficient as the K-means clustering. The best separations were obtained for three classes of grape skin anthocyanin.

Prediction Models for Assessing Lycopene in Open-Field Cultivated Tomatoes by Means of a Portable Reflectance Sensor: Cultivar and Growing-Season Effects.

Reflectance spectroscopy represents a useful tool for the nondestructive assessment of tomato lycopene, even in the field. For this reason, a compact, low-cost, light emitting diode-based sensor has been developed to measure reflectance in the 400-750 nm spectral range. It was calibrated against wet chemistry and evaluated by partial least squares (PLS) regression analyses. The lycopene prediction models were defined for two open-field cultivated red-tomato varieties: the processing oblong tomatoes of the cv. Calista (average weight: 76 g) and the fresh-consumption round tomatoes of the cv. Volna (average weight: 130 g), over a period of two consecutive years. The lycopene prediction models were dependent on both cultivar and season. The lycopene root mean square error of prediction produced by the 2014 single-cultivar calibrations validated on the 2015 samples was large (33 mg kg-1) in the Calista tomatoes and acceptable (9.5 mg kg-1) in the Volna tomatoes. A more general bicultivar and biyear model could still explain almost 80% of the predicted lycopene variance, with a relative error in red tomatoes of less than 20%. In 2016, the in-field applications of the multiseasonal prediction models, built with the 2014 and 2015 data, showed significant ( P < 0.001) differences in the average lycopene estimated in the crop on two sampling dates that were 20 days apart: on August 19 and September 7, 2016, the lycopene was 98.9 ± 9.3 and 92.2 ± 10.8 mg kg-1 FW for cv. Calista and 54.6 ± 13.2 and 60.8 ± 6.8 mg kg-1 FW for cv. Volna. The sensor was also able to monitor the temporal evolution of lycopene accumulation on the very same fruits attached to the plants. These results indicated that a simple, compact reflectance device and PLS analysis could provide adequately precise and robust (through-seasons) models for the nondestructive assessment of lycopene in whole tomatoes. This technique could guarantee tomatoes with the highest nutraceutical value from the production, during storage and distribution, and finally to consumers.

Prediction models for assessing anthocyanins in grape berries by fluorescence sensors: Dependence on cultivar, site and growing season.

Fluorescence sensors are useful tools for the non-destructive assessment of grape berry anthocyanins. The Multiplex (Mx) sensor here studied provides two anthocyanin indices: ANTHR = log(1/Chl-fluorescence_R) and ANTHRG = log(Chl-fluorescence_R/Chl-fluorescence_G), based on the chlorophyll (Chl) fluorescence excited with red (R) and green (G) light. These indices were calibrated against wet chemistry. The dependence of anthocyanin prediction models on cultivar, season and site was studied on four cultivars in two Italian regions during three consecutive years. The 2010 global model (all cultivars at both growing sites) gave relative prediction errors on anthocyanin content less than 14.1% (ANTHR) and 19.0% (ANTHRG). The ANTHRG was independent of season, maintaining a relative error of about 20% in both 2011 and 2012. In field applications of the calibrated Mx, it showed its ability to detect inter-plot and inter-season differences on both growing sites.

Nondestructive Optical Sensing of Flavonols and Chlorophyll in White Head Cabbage (Brassica oleracea L. var. capitata subvar. alba) Grown under Different Nitrogen Regimens.

A multiparametric optical sensor was used to nondestructively estimate phytochemical compounds in white cabbage leaves directly in the field. An experimental site of 1980 white cabbages (Brassica oleracea L. var. capitata subvar. alba), under different nitrogen (N) treatments, was mapped by measuring leaf transmittance and chlorophyll fluorescence screening in one leaf/cabbage head. The provided indices of flavonols (FLAV) and chlorophyll (CHL) displayed the opposite response to applied N rates, decreasing and increasing, respectively. The combined nitrogen balance index (NBI = CHL/FLAV) calculated was able to discriminate all of the plots under four N regimens (0, 100, 200, and 400 kg/ha) and was correlated with the leaf N content determined destructively. CHL and FLAV were properly calibrated against chlorophyll (R(2) = 0.945) and flavonol (R(2) = 0.932) leaf contents, respectively, by using a homographic fit function. The proposed optical sensing of cabbage crops can be used to estimate the N status of plants and perform precision fertilization to maintain acceptable crop yield levels and, additionally, to rapidly detect health-promoting flavonol antioxidants in Brassica plants.

Esculetin and esculin (esculetin 6-O-glucoside) occur as inclusions and are differentially distributed in the vacuole of palisade cells in Fraxinus ornus leaves: a fluorescence microscopy analysis.

The location of individual coumarins in leaves of Fraxinus ornus acclimated at full solar irradiance was estimated using their specific UV- and fluorescence spectral features. Using a combination of UV-induced fluorescence and blue light-induced fluorescence of tissues stained with diphenylborinic acid 2-amino-ethylester, in wide field or confocal laser scanning microscopy, we were able to visualize the distribution of esculetin and esculetin 6-O-glucoside (esculin) in palisade cells. Coumarins are not uniformly distributed in the cell vacuole, but accumulate mostly in the adaxial portion of palisade cells. Our study indeed shows, for the first time, that coumarins in palisade cells accumulate as vacuolar inclusions, as previously reported in the pertinent literature only for anthocyanins. Furthermore, esculetin and esculin have a different vacuolar distribution: esculetin largely predominates in the first 15 µm from the adaxial epidermis. This leads to hypothesize for esculetin and esculin different transport mechanisms from the endoplasmic reticulum to the vacuole as well as potentially different roles in photoprotection. Our study open to new experiments aimed at exploring the mechanisms that deliver coumarins to the vacuole using different fluorescence signatures of coumarin aglycones and coumarin glycosides.

Isoprene production in transgenic tobacco alters isoprenoid, non-structural carbohydrate and phenylpropanoid metabolism, and protects photosynthesis from drought stress.

Isoprene strengthens thylakoid membranes and scavenges stress-induced oxidative species. The idea that isoprene production might also influence isoprenoid and phenylpropanoid pathways under stress conditions was tested. We used transgenic tobacco to compare physiological and biochemical traits of isoprene-emitting (IE) and non-emitting (NE) plants exposed to severe drought and subsequent re-watering. Photosynthesis was less affected by drought in IE than in NE plants, and higher rates were also observed in IE than in NE plants recovering from drought. Isoprene emission was stimulated by mild drought. Under severe drought, isoprene emission declined, and levels of non-volatile isoprenoids, specifically de-epoxidated xanthophylls and abscisic acid (ABA), were higher in IE than in NE plants. Soluble sugars and phenylpropanoids were also higher in IE plants. After re-watering, IE plants maintained higher levels of metabolites, but isoprene emission was again higher than in unstressed plants. We suggest that isoprene production in transgenic tobacco triggered different responses, depending upon drought severity. Under drought, the observed trade-off between isoprene and non-volatile isoprenoids suggests that in IE plants isoprene acts as a short-term protectant, whereas non-volatile isoprenoids protect against severe, long-term damage. After drought, it is suggested that the capacity to emit isoprene might up-regulate production of non-volatile isoprenoids and phenylpropanoids, which may further protect IE leaves.

Potential of a multiparametric optical sensor for determining in situ the maturity components of red and white Vitis vinifera wine grapes.

A nondestructive fluorescence-based technique for evaluating Vitis vinifera L. grape maturity using a portable sensor (Multiplex) is presented. It provides indices of anthocyanins and chlorophyll in Cabernet Sauvignon, Merlot, and Sangiovese red grapes and of flavonols and chlorophyll in Vermentino white grapes. The good exponential relationship between the anthocyanin index and the actual anthocyanin content determined by wet chemistry was used to estimate grape anthocyanins from in field sensor data during ripening. Marked differences were found in the kinetics and the amount of anthocyanins between cultivars and between seasons. A sensor-driven mapping of the anthocyanin content in the grapes, expressed as g·kg(-1) fresh weight, was performed on a 7-ha vineyard planted with Sangiovese. In the Vermentino, the flavonol index was favorably correlated to the actual content of berry skin flavonols determined by means of HPLC analysis of skin extracts. It was used to make a nondestructive estimate of the evolution in the flavonol concentration in grape berry samplings. The chlorophyll index was inversely correlated in a linear manner to the total soluble solids (°Brix): it could, therefore, be used as a new index of technological maturity. The fluorescence sensor (Multiplex) possesses a high potential for representing an important innovative tool for controlling grape maturity in precision viticulture.